LS0302 Appendix
Figures , Publications and Technology Transfer
Figure 2.2.1 Diurnal variation in total VFA concentrations in Rusitec vessels fed different
alternative forage silages. Each value represents a mean of 3 replicated experiments.
Total VFA (mmol/l)
120.0
100.0
Barley/Pea
Red clover
80.0
Grass
60.0
Kale
Lotus
40.0
Lucerne
20.0
0.0
0
5
10
Time (h)
15
20
25
Figure 4.1.1 Relationship between the concentration of iso C15:0 () or iso C17:0 ()
and DM disappearance (g/g DM) in feed bolus residues of a) grass or b) white clover.
A
B
180
120
160
100
140
80
mg/kg DM
mg/kg DM
120
100
80
60
60
40
40
20
20
0
0
0.2
0.4
0.6
0.8
0
1
0
DM disappearance (g/g DM)
0.2
0.4
0.6
0.8
1
DM disappearance (g/g DM)
Figure 4.1.2 Polymerase chain reaction-denaturing gradient gel electrophoresis (PCRDGGE) analysis of Dacron bag residue incubated in the rumen for different times
Grass boli
M 0
Mrs
2
8
24
0 2 8 24 h
White clover boli
0 2 8 24 72 0 2 8 24 72
hrs
B. fragilis
R. albus SY3
S. bovis
E. coli
R. albus 7
R. albus 8
F. succinogenes
F. succinogenes
Cow A
Cow B
Cow C
Cow D
Figure 4.1.3 Anaerobic fungal populations detected in boli
incubated in the rumen of dairy cows
Boli from Grass
M 0 1 2
Neocallimastix
Anaeromyces
Piromyces
Orpinomyces
4 8 24 72 h
Boli from WhiteClover
M 0 1 2
4 8 24 72 h
a)
b)
1
1
Protein disappearance
(g/g initial DM)
DM disappearance (g/g DM)
Figure 4.2.1. In situ disappearances of a) dry matter (DM) and b) protein from grass () or white
clover () boli.
0.8
0.6
0.4
0.2
0
0
5
10
15
20
25
0.8
0.6
0.4
0.2
30
0
0
5
Incubation period (hrs)
10
15
20
25
30
Incubation period (hrs)
Incubation period (h)
Figure 4.2.2. SDS-PAGE separation of polypeptides extracted from a) grass and b) white clover
after incubation in the rumen. Ten g protein was loaded per track. Broad range molecular weight
markers (BioRad, UK) are indicated (M).
a) M
0
1
2
4
8
24
b) 0
1
2
4
8
24
A
B
C
Boli from grass
Boli from white clover
A = Large subunit Rubisco; B = Small subunit Rubisco; C = Light harvesting protein
Figure 4.2.3. Immuno-detection of the Rubisco large subunit (LSU) extracted from a) grass and b)
white clover after incubation in the rumen.
0
1
2
4
8
24
0
1
2
4
8
24
Rubisco
a)
Boli from grass
b)
Boli from white clover
Broad range molecular weight markers (BioRad, UK) are indicated in the left hand lane of the gel
Figure 4.3.1 Polypeptides in Lolium perenne before and after 16h in sacco incubation in
Dacron bags, polythene bags or dialysis tubing. Lp; grass before treatment, A-D;
herbage samples recovered from the rumen of four cows, molecular weight standards
are as indicated on right hand side of figure. Arrows show Rubisco large and small
subunits (LSU, SSU) and light harvesting protein (LHCPII). Gels loaded at 10 g
protein per track.
Figure 4.3.2 Quantification of polypeptide bands resolved by SDS-PAGE showing
proportions of LHCPII, and the Rubisco large and small subunits. Treatments were:
open bars, fresh grass; crossed bars, Dacron bags; cross-hatched bars, polythene bags;
solid bars, dialysis tubing.
75
Band density (arbitrary units)
Fresh grass
Dacron
60
Polythene
Dialysis
45
30
15
0
LSU
LHCPII
SSU
Figure 4.4.1 Disappearance of dry matter from the bolus, fresh and freeze-dried grass
after different times on incubation under in vitro conditions.
100
90
80
Dry matter remaining in the residue (%)
70
60
Bolus
Fresh Grass
50
Freeze Dried Grass
40
30
20
10
0
0
10
20
30
40
50
60
70
80
Time since inoculation (h)
Figure 4.4.2 Disappearance of water soluble carbohydrate from the bolus, fresh and
freeze-dried grass after different times of incubation under in vitro conditions.
100.00
90.00
Dry Matter Disappearance (%)
80.00
70.00
60.00
Bolus
50.00
Fresh Grass
Freeze Dried
Grass
40.00
30.00
20.00
10.00
0.00
0
10
20
30
40
Time since inoculation (h)
50
60
70
80
Publications
Refereed papers
Davies, D. R. , Merry, R. J. , Williams, A. P. , Bakewell, E. L. , Leemans, D. K. & Tweed,
J. K. S. (1998) Nutrition, feeding and calves. Proteolysis during ensilage of forages varying
in soluble sugar content Journal of Dairy Science 81 (2) 444-453. [LS0302)
Dewhurst, R. J. , Davies, D. R. & Merry, R. J. (2000). Microbial protein supply from the
rumen. Animal Feed Science and Technology 85 (1-2) 1-21 Elsevier Science. [LS0302]
Dewhurst, R.J., Fisher, W.J., Tweed, J.K.S. & Wilkins, R.J. Inoculated legume silages for
milk production. 1. Intake and milk production. Journal of Dairy Science. Submitted
[LS0302]
Dewhurst, R.J., Scollan, N.D., Moorby, J.M., Merry, R.J. & Wilkins, R.J. Inoculated
legume silages for milk production. 2. Effects on rumen function. Journal of Dairy Science.
submitted [LS0302]
Dewhurst, R. J. , Merry, R. J. , Bertilsson, J. & Tuori, M. (2002). Effects of legume
silages on diet digestibility and rumen function Legume Silages for Animal Production LEGSIL. Proceedings of an International Workshop, Braunschweig, Germany, 8-9 July 2001.
(Landbauforschung Volkenrode, Special Issue 234) 47-53. [LS0302]
Fraser, M. D. , Winters, A. L. , Fychan, A. R. , Davies, D. R. & Jones, R. (2001). The
effect of harvest date and inoculation on the yield, fermentation characteristics and feeding
value of kale silage Grass and Forage Science 56 (2) 151-161. [LS0301, 02]
Lee, M.R.F., Harris, L.J., Dewhurst, R.J., Merry, R.J. & Scollan, N.D. The effect of
clover silages on long chain fatty acid rumen transformations and digestion in beef steers.
Animal Science submitted [LS0302]
Merry, R. J. , Jones, R. & Theodorou, M. K. (2001) Alternative forages - back to the
future Biologist 48 (1) 30-34. [LS0301, 02]
Merry, R. J. , Jones, R. & Theodorou, M. K. (1999). The conservation of grass. Grass: Its
Production and Utilization Hopkins, A., eds. 196-228 (Chapter 8) Blackwell Science,
Oxford. [LS0301, 02]
Winters, A. , Lloyd, J. D. , Jones, R. & Merry, R. J. (2002). Evaluation of a rapid method
for estimating free amino acids in silages Animal Feed Science and Technology Vol 99, 177 –
187. [LS0301/02]
Winters, A. L. , Cockburn, J. E. , Dhanoa, M. S. & Merry, R. J. (2000). Effects of lactic
acid bacteria in inoculants on changes in amino acid composition during ensilage of sterile
and non-sterile ryegrass Journal of Applied Microbiology 89 (3) 442-451. [LS0302]
Reports
Jones, R. , Theodorou, M. K. , Merry, R. J. , Adesogan, A. T. , Davies, D. R. , Dewhurst,
R. J. et al. (2000) Conservation of alternative forage crops Final report to Milk Development
Council (MDC) - June 2000 IGER, Aberystwyth.
Abstracts
Akyol, I. , Fraser, M. D. , Griffith, G. W. & Davies, D. R. (1999). Comparison of in vivo
and in vitro digestibility of silages made from a range of alternative crops. Proceedings of
the British Society of Animal Science (BSAS) Annual Meeting, Scarborough, UK, 22-24
March 1999 97 BSAS, Penicuik, UK
Davies, D. R. , Leemans, D. K. & Merry, R. J. (2002). Ensiling either high or low sugar
containing perennial ryegrasses with or without red clover Grassland science in Europe
Poster Multi-Function Grasslands: Quality Forages, Animal Products and Landscapes.
Proceedings 19th General Meeting of the European Grassland Federation (EGF), La
Rochelle, France, 27-30 May 2002 Durand, J-L. , Emile, J-C. , Huyghe, C. , Lemaire, G., eds.
7 194-195.
Davies, D. R. , Leemans, D. K. & Merry, R. J. (2002). Improving silage quality by
ensiling perennial ryegrasses high in water soluble carbohydrate content either with or
without different additives. The XIIIth International Silage Conference SAC, Auchincruive,
Ayr, Scotland, 11-13 September 2002, p. 386.
Davies, D.R., Dhanoa, M.S., Leemans, D.K. & Merry, R.J. (2002). Relationships
between chemical indicators of silage quality and laboratory-measured aerobic spoilage. The
XIIIth International Silage Conference SAC, Auchincruive, Ayr, Scotland, 11-13 September
2002, p. 118.
Davies, D. R. , Brooks, A. E. , Dhanoa, M. S. , Merry, R. J. & Theodorou, M. K. (2000).
Development of novel approaches to examine rumen microbial succession Reproduction
Nutrition and Development 40, 181.
Davies, D.R., Leemans, D.K., Bakewell, E.L., Lowes, K., Winters, A.L., Scollan, N.D.,
Evans, P., Cooper, A., & Merry, R.J. (2002). Development of a novel silage inoculant
from small scale to large clamp silos. The XIIIth International Silage Conference SAC,
Auchincruive, Ayr, Scotland, 11-13 September 2002, p. 202.
Davies, D. R. , Winters, A. L. , Leemans, D. K. , Dhanoa, M. S. & Merry, R. J. (1999).
The effect of inoculant treatment of alternative crop forages on silage quality and in vitro
rumen function Silage Production in Relation to Animal Performance, Animal Health, Meat
and Milk Quality, Proceedings of the XIIth International Silage Conference, Uppsala,
Sweden, 5-7 July 1999 Pauly, T., eds. 131-132 (Poster abstracts) Swedish University of
Agricultural Sciences.
Dewhurst, R. J. , Merry, R. J. , Davies, D. R. , Moorby, J. M. , Scollan, N. D. , Neville,
M. A. et al. (2000). Effects of legume silages on the quantity and particle size distribution of
rumen contents in Holstein-Friesian cows Proceedings of the British Society of Animal
Science (BSAS) Annual Meeting, Scarborough, UK, 20-22 March 2000 1 BSAS.
Fychan, A. R. , Winters, A. , Lloyd, J. D. & Jones, R. (2000). The ensiling potential of red
clover / lucerne bi-crops Proceedings 6th British Grassland Society Research Conference,
Aberdeen, 11-13 September 2000 63-64.
Kim, E. J. , Dewhurst, R. J. , Merry, R. J. , Davies, D. R. & Theodorou, M. K.
Use of odd-chain fatty acid profiles to study microbial colonisation of boli from freshly
ingested herbages Abstracts Beyond antimicrobials - the future of gut microbiology. 3rd Joint
Symposium organised by INRA (France) and RRI (UK), Aberdeen, 12-15 June 2002.
Kim, E. J. , Evans, R. T. , Tweed, J. K. S. , Davies, D. R. , Merry, R. J. & Dewhurst, R.
J. (2002) Odd-chain fatty acids as markers of the microbial colonisation of freshly-ingested
grass and microbial contamination of dacron bag residues Abstract Proceedings of the British
Society of Animal Science (BSAS) Annual Conference, York, April 2002 British Society of
Animal Science, p. 50.
Lee, M. R. F. , Evans, R. T. , Dhanoa, M. S. , Merry, R. , Dewhurst, R. J. & Scollan, N.
D. (2002) Biohydrogenation and duodenal flow of C18 polyunsaturated fatty acids in steers
fed grass or grass:legume silages Abstract Proceedings of the British Society of Animal
Science (BSAS) Annual Conference, York, April 2002 British Society of Animal Science,
Penicuik, p. 10.
Lee, S. S. , Chang, M. B. , Scollan, N. D. , Merry, R. J. , Dhanoa, M. S. , Hobbs, P. J. et al.
(1999). The fatty acid composition of solid- and liquid-associated rumen bacteria isolated
from cows Proceedings of the British Society of Animal Science (BSAS) Annual Meeting,
Scarborough, UK, 22-24 March 1999 30 BSAS, Penicuik, UK.
Merry, R. J. , Leemans, D. K. & Davies, D. R. (2002). Improving the efficiency of silageN utilisation in the rumen through the use of perennial ryegrasses high in water-soluble
carbohydrate content. The XIIIth International Silage Conference SAC, Auchincruive, Ayr,
Scotland, 11-13 September 2002, p. 374.
Minchin, F. R. , Winters, A. , Kingston-Smith, A. H. & Robbins, M. P. (2000). Can
protein losses be reduced during the conversion of legume protein into animal protein? Plant
Form and Function - Adaptations to Stress. Abstracts of a Meeting, University of Western
Australia, Nedlands, 27-29 September 2000 45-47.
Speijers, M. H. M. , Fraser, M. D. , Fychan, R. , Theobald, V. J. & Winters, A. (2002).
Evaluation of different silage additives for ensiling lucerne and red clover. Proceedings XIII
International Silage Conference, Auchincruive, Scotland, 11-13 September 2002 Gechie, L.
M. , Thomas, C., eds. 112-113.
Winters, A.L. & Mincin, F.R. (2002). The effect of PPO on the protein content of ensiled
red clover. The XIIIth International Silage Conference SAC, Auchincruive, Ayr, Scotland, 1113 September 2002, p. 84.
Winters, A. , Lloyd, J. , Leemans, D. , Bakewell, E. & Merry, R. (2000). Effect of
Lactobacillus plantarum on protein degradation during ensilage of red clover Proceedings 6th
British Grassland Society Research Conference, Aberdeen, 11-13 September 2000 33-34
British Grassland Society, Reading.
Winters, A. L. , Fychan, A. R. , Davies, D. R. , Lloyd, J. D. , Merry, R. J. & Jones, R.
(1999). Protein content of a range of ensiled legumes Silage Production in Relation to Animal
Performance, Animal Health, Meat and Milk Quality, Proceedings of the XIIth International
Silage Conference, Uppsala, Sweden, 5-7 July 1999 Pauly, T., eds. 170-171 (Poster abstracts)
Swedish University of Agricultural Sciences.
Winters, A. L. , Lloyd, J. D. , Lowes, K. F. , Jones, R. & Merry, R. J. (1999). A rapid and
economical technique for predicting free amino acid and content in legume silage. Silage
Production in Relation to Animal Performance, Animal Health, Meat and Milk Quality,
Proceedings of the XIIth International Silage Conference, Uppsala, Sweden, 5-7 July 1999
Pauly, T., eds. 168-169 (Poster abstracts) Swedish University of Agricultural Sciences.
Winters, A. L. , Lloyd, J. D. , Fychan, A. R. , Davies, D. R. , Merry, R. J. & Jones, R.
(1998) Protein degradation during ensilage of red clover and lucerne forage. Keeping the
Balance, British Grassland Society (BGS) Winter Meeting, Peebles, Scotland, UK, 30
November - 1 December 1998 18.1-18.2.
Winters, A.L., Davies, Z., Minchin, F.R. & Merry, R.J., (2002). Effect of protein content
of white clover on silage quality. The XIIIth International Silage Conference SAC,
Auchincruive, Ayr, Scotland, 11-13 September 2002, p. 106.
Winters, A.L., Lloyd, J., Leemans, D.K. Lowes, K. & Merry, R.J., (2002). Effect of
inoculation with Lactobacillus plantarum on protein degradation during ensilage of red
clover. The XIIIth International Silage Conference SAC, Auchincruive, Ayr, Scotland, 11-13
September 2002, p. 108.
Popular articles
Davies, D. R. (1999). Making the most of home fodder Farmers Weekly (19/11/99) S18.
Davies, D. R. (2002). Farmers urged to make more of grass sugars. Y Tir & Welsh Farmer
(June) 3.
Davies, D. R. & Merry, R. J. (2001). Sweet grass, sweeter silage IGER Press Release.
Technical bulletins
Davies, D. R. , Merry, R. J. & Leemans, D. K. (2000). Silage inoculants - making the most
of grass sugars IGER Information leaflet. (Published July 2000) IGER, Aberystwyth, 2 pp.
Technology Transfer
01/11/2000 am
Farmers Presentation
01/11/2000 pm
Local British Grassland Society Meeting
01/12/99
Local British Grassland Society Meeting
01/12/99 am
Farmers Presentation
01/12/99 am
Farmers Presentation
01/12/99 am
Farmers Presentation
01/2/2000 am
Farmers Presentation
01/2/2000 pm
Farmers Presentation
02/11/2000 am
02/2/2000 am
Farmers Presentation
Farmers Presentation
02/2/2000 pm
03/2/2000 am
Farmers Presentation
Farmers Presentation
03/3/2000 pm
03/8/2000
06/11/2000 pm
06/11/2001
06/12/1999 pm
06/2/2001
07/11/2000 am
07/11/2000 pm
07/12/1999 am
07/12/1999 pm
08/12/1999 am
Farmers Presentation
Farmers Presentation
Farmers Presentation
Presentation – ADAS - IGER MOU
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
08/12/1999 pm
09/12/1999 am
Farmers Presentation
Farmers Presentation
10/1/2001
10/3/1999
11/7/2000
12/12/2000 pm
13/11/2000 am
16/1/2001
17/1/2001
18/1/2001 am
18/1/2001 pm
18/7/2000
19/10/1999
21/11/2000 pm
22/11/2000
22/11/1999
Presentation to Promar Consultants
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Presentation to Farm Advisors
Local British Grassland Society Meeting
Farmers Presentation
Farmers Presentation
29/11/1999 pm
29/12/1999 am
Farmers Presentation
Farmers Presentation
30/11/1999 am
30/11/1999 pm
31/10/2000 pm
31/11/1999 am
31/11/2999 pm
9/3/1999
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Farmers Presentation
Glastonbury, Somerset
Somerset
Gatwick
Ayr, Ayrshire
Lanark, Lanarkshire
Winchester, Wiltshire
Ballymena Road, Co.
Antrim
Coleraine, Co.
Londonderry
Devon
Fivemiletown, Co.
Tyrone
Armagh, Co. Armagh
Ballynahinch, Co.
Down
Cookstown, Co. Tyrone
Newtown
Uttoxeter, Staffs
IGER
St. Asaph, Denbighshire
Reaseheath
Derbyshire
Leicestershire
Wigton, Cumbria
Gisbon Lancashire
Driffield, West
Yorkshire
Thirsk, North Yorkshire
Dumfries, Dumfries &
Galloway
Bristol
Carlisle
IGER
Carlisle, Cumbria
Lleyn, Gwynedd
Compton
Carlisle
Penrith
Lancaster Market
IGER
IGER
Isle of Wight
Dorset
Clynderwen,
Carmarthenshire
Glastonbury, Somerset
Gloucester,
Gloucestershire
Barnstaple, Devon
Bodmin, Cornwall
Romsey, Hampshire
Exeter, Devon
Shaftesbury, Dorset
Preston
IGER Stand at the Pembrokeshire Show, Haverfordwest 15th August 2000
IGER Stand at the Royal Show, Stoneleigh 4th- 5th July 2000
IGER Stand at the Royal Welsh Show, Builth Wells 24th- 27th July 2000